Effects of Mechanical Stress and Phenotype Switching on Human Stem Cell-Derived Vascular Smooth Muscle Cells: Modeling Gene Regulatory Networks

该项目将揭示人类血管平滑肌细胞(VSMCs)如何适应血压等机械力，从有利于形成新血管的状态转变为控制流向组织的血液的状态。尽管这一转变在人类血管发育和组织修复中的重要性，但对人类这种转变的机制了解甚少。除了科学进步，该项目还将促进本科生和研究生在细胞生物学、物理学和计算建模方面的培训和教育，为他们在学术界和工业界的职业生涯做好准备。当地一所少数民族占多数的磁铁数学科学高中的研究实习也将为巴尔的摩市的学生提供培训机会，以鼓励以STEM为基础的职业。从长远来看，对社会的好处可能包括新的方法来模拟或利用vSMC进行血管形成和血管修复。本研究的目标是利用围产期发育的人类vSMC替代模型来确定机械力如何调节vSMC表型转换。这里使用的模型系统依赖于从具有明确的合成和收缩表型状态的人诱导的多能干细胞分化而来的谱系特异性vSMCs。这些VSMC的机械性能和对应力/应变的响应将使用能够在2D和3D几何形状中进行单细胞分析和拉伸的平台来评估。一旦“调整”以最大限度地发挥内力，模型系统将与RNA-seq、生物信息学和计算建模一起使用，以确定关键调控因子(例如，转录因子、信号分子)，并对预计将控制hPSC-vSMC对增加的压力/应变的反应的调控网络进行建模。结合CHIP-SEQ分析，结果将被用于破译机械力如何影响导致vSMC表型转变的转录机制。该项目由分子和细胞生物科学部的细胞动力学和功能(CDF)以及系统和合成生物学(SSB)集群联合资助。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

This project will reveal how human vascular smooth muscle cells (vSMCs) adapt to mechanical forces like blood pressure to switch from a state that favors the formation of new vessels to one designed to control blood flow to tissues. The mechanisms responsible for this transition in humans are poorly understood, despite the importance of this switch in human vascular development and tissue repair. Beyond scientific advances, the project will promote the training and education of undergraduate and graduate students in cell biology, physics and computational modeling that will prepare them for careers in academia and industry. Research internships with a local majority-minority magnet math-science high school will also offer training opportunities for Baltimore City students to encourage STEM-based careers. Long-term the benefits to society will likely include new approaches to model or utilize vSMCs for vessel formation and vascular repair.The goal of this research is to determine how mechanical forces regulate vSMC phenotype switching using a human vSMC surrogate model of perinatal development. The model system employed here relies on lineage-specific vSMCs differentiated from human induced pluripotent stem cells with well-defined synthetic and contractile phenotypic states. The mechanical properties and response of these vSMCs to stress/strain will be assessed using platforms that enable single cell analyses and stretching of cells in both 2D and 3D geometries. Once “tuned” to maximize internal force development, the model systems will be used with RNA-seq, bioinformatics, and computational modeling to identify key regulatory factors (e.g., transcription factors, signaling molecules) and to model regulatory networks predicted to control hPSC-vSMC responses to increased stress/strain. When coupled with ChIP-seq assays, the results will be used to decipher how mechanical forces affect transcriptional mechanisms responsible for phenotype transitions in vSMCs.This project is funded jointly by the Cellular Dynamics and Function (CDF) and Systems and Synthetic Biology (SSB) clusters of the Division of Molecular and Cellular Biosciences.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.